Simplifying large-scale sequencing projects, such as genome-scale projects, depends on rapid acquisition of DNA sequence data. Unfortunately, nucleic acid sequencing procedures typically involve time-consuming cycling of reagents, enzymatic reactions that must go to completion, and potentially long periods of acquiring measurement data. Of course, all of this is a legacy of the early nucleic acid sequencing paradigms that relied on processing one nucleotide position at a time.
Alternative methods aimed at identifying short stretches of sequence at one time (e.g., using sequencing-by-hybridization) have not found wide acceptance for determining unknown sequences. Instead, these techniques generally are applied to investigating subtle changes in sequences that already are known. The same is true for techniques that are based on “melting curve analysis,” where the interrogation sequences are commonly no larger than about twenty nucleotides in length.
The techniques described herein provide an approach that advantageously speeds acquisition of nucleic acid sequence data and provide other advantages as well.